US 12,435,340 B2
Methods for transformation of dicot plant cells
Jianping Xu, Beijing (CN); Chao Li, Beijing (CN); and Qin Wen, Beijing (CN)
Assigned to Syngenta Crop Protection AG, Basel (CH)
Appl. No. 17/640,099
Filed by SYNGENTA CROP PROTECTION AG, Basel (CH)
PCT Filed Aug. 28, 2020, PCT No. PCT/US2020/048308
§ 371(c)(1), (2) Date Mar. 3, 2022,
PCT Pub. No. WO2021/045960, PCT Pub. Date Mar. 11, 2021.
Claims priority of application No. PCT/CN2019/104384 (WO), filed on Sep. 4, 2019.
Prior Publication US 2022/0290168 A1, Sep. 15, 2022
Int. Cl. C12N 15/82 (2006.01)
CPC C12N 15/8205 (2013.01) [C12N 15/8207 (2013.01)] 16 Claims
 
1. A method of transforming a tomato plant cell, comprising:
(a) obtaining a tomato plant seed;
(b) soaking the tomato plant seed in a seed-soak medium;
(c) dividing the soaked tomato plant seed of step (b) into at least 10 tomato explants;
(d) inoculating the at least 10 tomato explants of step (c) with Agrobacterium comprising a nucleic acid of interest, thereby obtaining Agrobacterium-inoculated tomato explants; and
(e) co-cultivating the Agrobacterium-inoculated tomato explants to produce a transformed tomato plant cell, wherein the soaking step of step (b) is within the range of 1 hour to 72 hours, with or without light, wherein the method produces less than 15% aberrant tetraploid transformants, wherein the plurality of tomato explants is at least 10 discrete explants.